Comparison of Detection Methods for <i>Legionella</i> Species in Environmental Water by Colony Isolation, Fluorescent Antibody Staining, and Polymerase Chain Reaction

Yamamoto, Hiroyuki; Hashimoto, Yasuhiro; Ezaki, Takayuki

doi:10.1111/j.1348-0421.1993.tb01684.x

Cited by 57 publications

(46 citation statements)

References 21 publications

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“…Furthermore, the methodological problems in the isolation of legionellae from environmental water can also contribute to an underestimation of culturable Legionella cells. The use of a selective medium and pretreat- ment by acid or heating reduce the cultivability (58). For all these reasons, it could be concluded that the culture methods underestimate the population of Legionella.…”

Section: Discussionmentioning

confidence: 99%

“…Molecular methods have also previously been developed to increase the rapidity of Legionella analysis. They are able to achieve a high degree of sensitivity and specificity without the need for a complex cultivation and additional confirmation steps (6,28,31,32,41,49,52,54,58). Some of these methods allow the detection of culturable and/or nonculturable Legionella within hours instead of the days required with the reference culture method.…”

Section: Discussionmentioning

confidence: 99%

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Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Delgado‐Viscogliosi

Simonart

Parent

et al. 2005

Appl Environ Microbiol

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A sensitive and specific method has been developed to enumerate viable L. pneumophila and other Legionella spp. in water by epifluorescence microscopy in a short period of time (a few hours). This method allows the quantification of L. pneumophila or other Legionella spp. as well as the discrimination between viable and nonviable Legionella. It simultaneously combines the specific detection of Legionella cells using antibodies and a bacterial viability marker (ChemChrome V6), the enumeration being achieved by epifluorescence microscopy. The performance of this immunological double-staining (IDS) method was investigated in 38 natural filterable water samples from different aquatic sources, and the viable Legionella counts were compared with those obtained by the standard culture method. The recovery rate of the IDS method is similar to, or higher than, that of the conventional culture method. Under our experimental conditions, the limit of detection of the IDS method was <176 Legionella cells per liter. The examination of several samples in duplicates for the presence of L. pneumophila and other Legionella spp. indicated that the IDS method exhibits an excellent intralaboratory reproducibility, better than that of the standard culture method. This immunological approach allows rapid measurements in emergency situations, such as monitoring the efficacy of disinfection shock treatments. Although its field of application is as yet limited to filterable waters, the double-staining method may be an interesting alternative (not equivalent) to the conventional standard culture methods for enumerating viable Legionella when rapid detection is required.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Delgado‐Viscogliosi

Simonart

Parent

et al. 2005

Appl Environ Microbiol

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“…A number of PCR-based assays have been developed for the detection and quantification of Legionella in water mainly using the 5S and 16S rRNA genes and the macrophage infectivity potentiator (mip) gene of L. pneumophila [23][24][25][26][27][28][29][30][31]. However, these assays lack the ability to discriminate between living and dead (non-infectious) Legionella cells.…”

Section: Introductionmentioning

confidence: 99%

Identification of Legionella spp. in Environmental Water Samples by ScanVIT-Legionella™ Method in Spain

Grúas

Alvarez²,

Lara

et al. 2013

Indian J Microbiol

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Rapid and more sensitive methods for the detection and quantification of viable Legionella cells have been developed. In this paper, a comparative analysis of environmental water samples using the ScanVIT-Legionella TM method and the traditional ''gold standard'' method of culturing is realised indicating the usefulness of the ScanVIT method. The ScanVIT-Legionella TM method was performed on environmental water samples from different locations of Huesca region (Spain). Legionella micro-colonies should appear green colour and Legionella pneumophila micro-colonies appear red. Twenty-one environmental water samples analysed by standard culture plus five control samples (Two sterile water samples with Legionella as positive controls and three sterile water samples as negative controls). All of them were used to apply ScanVIT-Legionella TM method. From of 21 environmental samples eleven were positive, six negative with both methods and four samples were negative for culture method and positive for ScanVIT-Legionella TM method. The positive control samples were positive and the negative were negative for both methods. A comparative analysis of the results obtained with two methods showed a strong positive determination coefficient (R 2 = 0.99753). The results demonstrate the usefulness of the ScanVITLegionella TM method as a rapid diagnostic tool in order to provide a diagnosis as quick as possible. ScanVITLegionella TM method offers a series of advantages such as quickly diagnosis, higher sensitivity and the possibility to identify Legionella spp. and L. pneumophila simultaneously.

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“…These two methods cannot detect non-L. pneumophila spp. Although L. pneumophila is the most frequent cause of legionellosis, non-L. pneumophila species also may cause serious or fatal disease (16,24). Non-L. pneumophila species in respiratory specimens are not detectable by the L. pneumophila direct fluorescent antigen test.…”

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confidence: 99%

Two-Step Scheme for Rapid Identification and Differentiation of Legionella pneumophila and Non- Legionella pneumophila Species

Zhan

et al. 2010

J Clin Microbiol

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A rapid two-step scheme based on PCR amplification and enzymatic digestion analysis of a 226-bp fragment of the 16S rRNA gene was developed to identify the Legionella genus by PCR amplification and to differentiate the Legionella pneumophila and non-Legionella pneumophila species by enzymatic digestion analysis. Among 42 ATCC strains (16 strains of L. pneumophila and 26 strains of non-L. pneumophila) and 200 Legionella isolates from environmental water samples, including pools, rivers, lakes, and cooling towers in Guangdong province, 99.59% of L. pneumophila and non-L. pneumophila strains were correctly identified and differentiated by this scheme. The procedure of this two-step identification and differentiation scheme is simple and takes only about 4 h. These results suggest that this two-step scheme provides a simple and convenient method for the rapid identification and differentiation of L. pneumophila and non-L. pneumophila species.

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Comparison of Detection Methods for Legionella Species in Environmental Water by Colony Isolation, Fluorescent Antibody Staining, and Polymerase Chain Reaction

Cited by 57 publications

References 21 publications

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Rapid Method for Enumeration of Viable Legionella pneumophila and Other Legionella spp. in Water

Identification of Legionella spp. in Environmental Water Samples by ScanVIT-Legionella™ Method in Spain

Two-Step Scheme for Rapid Identification and Differentiation of Legionella pneumophila and Non- Legionella pneumophila Species

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